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Gut Content Analysis of a Phloem-Feeding Insect, Bactericera cockerelli (Hemiptera: Triozidae)

W. Rodney Cooper, David R. Horton, Thomas R. Unruh, Stephen F. Garczynski
Environmental entomology 2016 v.45 no.4 pp. 938-944
Bactericera cockerelli, Candidatus Liberibacter solanacearum, DNA, Solanum dulcamara, Solanum tuberosum, Triozidae, chloroplasts, digestive system, genes, host plants, insects, internal transcribed spacers, landscape ecology, melting, pathogens, pests, potatoes, quantitative polymerase chain reaction, rearing, restriction endonucleases, sequence analysis, zebra chip disease
Potato psyllid, Bactericera cockerelli (Šulc) (Hemiptera: Triozidae), is a key pest of potato (Solanum tuberosum L., Solanales: Solanaceae) and a vector of “Candidatus Liberibacter solanacearum,” the pathogen associated with zebra chip disease. In addition to its presence on cultivated crops, the psyllid regularly occurs on numerous uncultivated annual and perennial species within the Solanaceae. A better understanding of landscape-level ecology of B. cockerelli would substantially improve our ability to predict which potato fields are most likely to be colonized by infected psyllids. We developed three PCR-based methods of gut content analysis to identify what plant species B. cockerelli had previously fed upon. These methods included—1) sequencing PCR amplicons of regions of plant-derived internal transcribed spacer (ITS) or the chloroplast trnL gene from psyllids, 2) high-resolution melting analysis of ITS or trnL real-time PCR products, and 3) restriction enzyme digestion of trnL PCR product. Each method was used to test whether we could identify psyllids that had been reared continuously on potato versus psyllids reared continuously on the perennial nightshade, Solanum dulcamara. All three methods of gut content analysis correctly identified psyllids from potato and psyllids from S. dulcamara. Our study is the first to demonstrate that plant DNA can be detected in a phloem-feeding insect. Gut content analysis, in combination with other landscape ecology approaches, could help elucidate patterns in landscape-level movements and host plant associations of B. cockerelli.